scholarly journals Using the Spleen as an In Vivo Systemic Immune Barometer Alongside Osteosarcoma Disease Progression and Immunotherapy with α-PD-L1

Sarcoma ◽  
2018 ◽  
Vol 2018 ◽  
pp. 1-13 ◽  
Author(s):  
Justin E. Markel ◽  
Jabeen Noore ◽  
Eric J. Emery ◽  
Harley J. Bobnar ◽  
Eugenie S. Kleinerman ◽  
...  
Keyword(s):  
Disease Progression ◽  
T Regulatory Cells ◽  
Immune Cell ◽  
Cytotoxic T Cells ◽  
M2 Macrophage ◽  
Regulatory Cells ◽  
Status Monitoring ◽  
Tumor Bearing ◽  
Marker Expression

Indications for immunotherapies are still unclear, and there is a great need for real-time patient immune status monitoring. In this study, we confirmed that the local and systemic immune profiles of an orthotopic osteosarcoma model with or without luciferase transfection were statistically equivalent. Next, we used flow cytometry to describe systemic immune cell populations influenced by osteosarcoma disease progression. When compared to vehicle-inoculated sham mice, it was found that tumor-bearing mice had significant immunophenotype disturbances at approximately 11 weeks after inoculation (at which time 90% of primary tumor-bearing mice have fulminant pulmonary metastases). Percent populations of natural killer cells and T regulatory cells were increased in the spleens of tumor-bearing mice (p<0.0083) compared to shams. Additionally, T lymphocytes from spleens of tumor-bearing mice showed increased Tim-3/PD-1 exhaustion status (p<0.0083). There were also increases in the percent populations of myeloid cells and overall M1/M2 macrophage marker expression on tumor-bearing mice spleens versus controls (p<0.00714). Finally, treatment with 20 μg α-PD-L1 decreased T-cell exhaustion back to sham status, with a corresponding increase in CTLA-4 expression on cytotoxic T cells in the majority of mice tested. Checkpoint inhibition also increased splenic monocyte maturation and returned macrophage M1/M2 marker expression back to sham status. These data suggest that cancer induces systemic immune dysregulation and that these changes may be elucidated and utilized for treatment purposes by sampling the systemic immune environment via the spleen. In addition, treatment with the checkpoint inhibitor α-PD-L1 may neutralize but not overcome the systemic immunological changes induced by a progressing malignancy.

scholarly journals Expansion of myeloid derived suppressor cells correlates with number of T regulatory cells and disease progression in myelodysplastic syndrome

2015 ◽  
Vol 5 (2) ◽  
pp. e1062208 ◽  
Author(s):  
Astrid Olsnes Kittang ◽  
Shahram Kordasti ◽  
Kristoffer Evebø Sand ◽  
Benedetta Costantini ◽  
Anne Marijn Kramer ◽  
...  
Keyword(s):  
Myelodysplastic Syndrome ◽  
Disease Progression ◽  
T Regulatory Cells ◽  
Suppressor Cells ◽  
Myeloid Derived Suppressor Cells ◽  
Regulatory Cells

Abstract 44: Failure of Protective Autoimmunity in Mouse and Human Atherosclerosis

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Dennis Wolf ◽  
Teresa Gerhardt ◽  
Nathaly Anto Michel ◽  
Bjarke Hansen ◽  
Alessandro Sette ◽  
...  
Keyword(s):  
T Cells ◽  
Lymph Nodes ◽  
Mhc Class Ii ◽  
T Regulatory Cells ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
T Cell Response ◽  
Regulatory Cells ◽  
Protective Autoimmunity

Background: In atherosclerosis, CD4 + T helper cells recognize auto-antigens including ApoB, the main protein in low-density lipoprotein (LDL). However, atherosclerosis-specific, auto-reactive CD4 + T cells have not been detected in vivo , and their function is unknown. Methods and Results: We have previously identified peptides derived from mouse ApoB that bind with high affinity to the MHC class II molecule of C57BL/6 mice (I-A b ). We designed and validated a new multimer of a recombinant MHC-II molecule fused to one ApoB auto-epitopes, P6 (TGAYSNASSTESASY, P6:I-A b ), that enabled detection of low-affinity, P6-reactive CD4 + T cells. Using this P6:I-A b multimer, we identified ApoB-reactive CD4 + T cells in healthy, young C57BL/6 mice that were predominately differentiated T-regulatory cells (T regs ) and expressed IL-10, a known atheroprotective cytokine. This population was detectable in lymph nodes and already showed a memory phenotype in young animals without atherosclerosis. In Apoe -/- mice, adoptively transferred ApoB P6-specific T regs accumulated in the aorta and draining lymph nodes and gave rise to pathogenic T H 1 and T H 17 cells. This phenotypic switch was caused by enhanced plasticity of antigen-specific T regs as evidenced by multiple clusters of intermediate T reg -T eff phenotypes in single cell RNA sequencing of 4485 antigen-specific CD4 + T cells. In the plaque, many T cells were ex-T regs as identified by a FoxP3 lineage tracker mouse, suggesting that atherosclerosis-specific CD4 + T cells lost their regulatory capacity. Vaccination with P6 maintained a protective phenotype in antigen-specific T regs and protected from atherosclerosis. In humans, ApoB-specific CD4 + T cells from atherosclerotic patients showed the same cytokine patterns found in mouse CD4 + T cells, suggesting that autoimmunity to ApoB is protective first, but later gives rise to a pathogenic CD4 + T cell response that aggravates atherosclerosis. Conclusion: Protective T-regulatory cells recognizing peptide antigens of ApoB exist in naïve mice, protect against atherosclerosis, but convert into pathogenic T H 1 and -17 cells during the natural course of disease in mice and humans. These results call for immunomodulatory therapies to maintain protective autoimmunity.

Immunosuppressive and metabolic agents that influence allo‐ and xenograft survival by in vivo expansion of T regulatory cells

2020 ◽  
Vol 27 (6) ◽  
Author(s):  
Yanli Zhao ◽  
Wenjun Hu ◽  
Pengfei Chen ◽  
Mengtao Cao ◽  
Yingwei Zhang ◽  
...  
Keyword(s):  
T Regulatory Cells ◽  
Regulatory Cells

scholarly journals Nanomedicine-mediated induction of immunogenic cell death and prevention of PD-L1 overexpression for enhanced hepatocellular carcinoma therapy

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Hanzhang Zhu ◽  
Weijiang Zhou ◽  
Yafeng Wan ◽  
Ke Ge ◽  
Jun Lu ◽  
...  
Keyword(s):  
Animal Model ◽  
Cell Death ◽  
Cytotoxic T Cells ◽  
Immunogenic Cell Death ◽  
Protein Levels ◽  
Tumor Bearing ◽  
Transmission Electron ◽  
Tumor Tissues ◽  
Programmed Death

Abstract Background The present study aims to develop a nanoparticle encapsulating doxorubicin (DOX) and programmed death-ligand 1 (PD-L1) siRNA and evaluate its anti-tumor effects on hepatoma carcinoma (HCC). Methods Nanoparticle encapsulating DOX and PD-L1 siRNA (NPDOX/siPD-L1) was characterized by dynamic light scattering and transmission electron microscopy. Flow cytometry was applied to analyze cell populations, NPDOX/siPD-L1 internalization, and cell apoptosis. Real-Time (RT)-quantitative reverse transcription (qPCR) and western blotting were used to determine the mRNA and protein levels, respectively. Released ATP was determined using ATP determination kit and cytokines were determined using specific ELISAs. A tumor-bearing animal model was established to evaluate the anti-tumor effects of NPDOX/siPD-L1. Results Treatment of NPDOX/siPD-L1 induced immunogenic cell death (ICD) and PD-L1 overexpression in HCC. In vivo study demonstrated that intravenously injection of NPDOX/siPD-L1 significantly inhibited the tumor volume and PD-L1 expressions of tumor tissue in the H22 tumor-bearing animal model. Besides, the treatment of NPDOX/siPD-L1 also regulated the populations of matured dendritic cells and cytotoxic T cells and the productions of cytokines in the tumor tissues. Conclusion Taken together, NPDOX/siPD-L1 showed significant anti-tumor effects on HCC by the induction of ICD and inhibition of PD-L1 overexpression.

scholarly journals Knockdown of microRNA-135b in Mammary Carcinoma by Targeted Nanodiamonds: Potentials and Pitfalls of In Vivo Applications

Nanomaterials ◽  
2019 ◽  
Vol 9 (6) ◽  
pp. 866 ◽  
Author(s):  
Romana Křivohlavá ◽  
Eva Neuhӧferová ◽  
Katrine Q. Jakobsen ◽  
Veronika Benson
Keyword(s):  
Antisense Rna ◽  
Tumor Tissue ◽  
Immune Cell ◽  
Ex Vivo ◽  
Tumor Bearing ◽  
Intravenous Application ◽  
Effective Inhibition ◽  
Tumor Accumulation ◽  
Safe Approach

Nanodiamonds (ND) serve as RNA carriers with potential for in vivo application. ND coatings and their administration strategy significantly change their fate, toxicity, and effectivity within a multicellular system. Our goal was to develop multiple ND coating for effective RNA delivery in vivo. Our final complex (NDA135b) consisted of ND, polymer, antisense RNA, and transferrin. We aimed (i) to assess if a tumor-specific coating promotes NDA135b tumor accumulation and effective inhibition of oncogenic microRNA-135b and (ii) to outline off-targets and immune cell interactions. First, we tested NDA135b toxicity and effectivity in tumorospheres co-cultured with immune cells ex vivo. We found NDA135b to target tumor cells, but it binds also to granulocytes. Then, we followed with NDA135b intravenous and intratumoral applications in tumor-bearing animals in vivo. Application of NDA135b in vivo led to the effective knockdown of microRNA-135b in tumor tissue regardless administration. Only intravenous application resulted in NDA135b circulation in peripheral blood and urine and the decreased granularity of splenocytes. Our data show that localized intratumoral application of NDA135b represents a suitable and safe approach for in vivo application of nanodiamond-based constructs. Systemic intravenous application led to an interaction of NDA135b with bio-interface, and needs further examination regarding its safety.

Expanding Tregs with IVIg

Blood ◽  
2008 ◽  
Vol 111 (2) ◽  
pp. 481-482 ◽  
Author(s):  
Rachel R. Caspi
Keyword(s):  
Autoimmune Diseases ◽  
Mechanism Of Action ◽  
T Regulatory Cells ◽  
Regulatory Cells

In this issue of Blood, Ephrem et al demonstrate that IVIg expands CD4+CD25+FoxP3+ T regulatory cells (Tregs) and enhances their function in vivo and in vitro. Their findings shed new light on the elusive mechanism of action of IVIg in ameliorating autoimmune diseases.

CD4+CD25+ T Cells Can Inhibit CD8 T Cell Mediated GVHD: Requirement for In Vivo Recognition of Allogeneic Host MHC Class II Antigens.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1307-1307
Author(s):  
Robert B. Levy ◽  
Angela Jones
Keyword(s):  
Weight Loss ◽  
T Cells ◽  
T Cell ◽  
Mhc Class Ii ◽  
Cd8 T Cells ◽  
T Regulatory Cells ◽  
Cd8 T Cell ◽  
Class Ii ◽  
Regulatory Cells

Abstract CD4 regulatory T (Treg) cells have shown promise in the transplantation mileu including the ability to inhibit the development of graft vs host disease (GVHD) following allogeneic hematopoietic stem cell transplants (HCT). The antigen specificity of the Treg population(s) involved is not yet clear nor is the role of their activation following transplant. We are interested in determining the requirement for recognition of host MHC antigens following infusion of CD4+CD25+ T cells in an experimental model of GVHD. To clearly distinguish the requirements of regulatory vs GVH reactive cells, a model of CD8 T cell mediated GVHD was developed using highly purified BALB/c (H2d) donor CD8+ T cells (Miltenyi column, 95-98%). CD8 T cells were transplanted together with T cell depleted (TCD) BALB/c BMC into 12.0 GY (6.0 Gy split dose) TBI conditioned C57BL/6 (B6, H2b) recipients. To support development of GVHD by these cells, resistance was inhibited by treatment of recipients with anti-NK1.1mab (PK136) at Days -1, 0 and +7. BALB/c CD8+ T cells at doses of 5.0x106 but not 2.5x106 induced weight loss and some lethality in B6 recipients. 5x106 CD8+ T cells were then transplanted into B6-MHC class II−/ − recipients. GVHD symptoms including weight loss and lethality were readily apparent in these mice post-transplant. Interestingly, GVHD was consistently more severe with respect to the induction of weight loss and lethality in MHC Class II−/ − vs B6-wt recipients. Highly enriched BALB/c CD4+CD25+ T cells (&gt; 95%) were produced from spleen and lymph node cells following negative (B-cells, CD8 and NK) and positive (CD25) selection using Miltenyi magnetic bead columns. Co-transplant of 1x106 CD4+CD25+ T cells together with BALB/c CD8+ T cells into B6 recipients inhibited GVHD as assessed by the absence of weight loss and lethality compared to B6 recipients of CD8+ T cells alone. In contrast, BALB/c CD4+CD25+ T cells failed to protect B6-MHC class II−/ − recipients from severe CD8+ T cell mediated GVHD. These findings demonstrate that donor CD4+ T regulatory cells can suppress GVHD inducing CD8+ T cells after the former recognize host class II alloantigen following transplant. We hypothesize that activated CD4+CD25+ T regulatory cells inhibit GVH reactive T cells at the host APC interface. Future studies in this model can be designed to examine ex-vivo activated and expanded CD4+CD25+ T regulatory populations. Transplant of such cells will enable us to address questions regarding the importance of in vivo recognition of host class II in the regulation of GVHD by these cells.

B Cell Depletion Reduces the Number and Function of T- Regulatory Cells and Enhances the Anti Tumor response

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3668-3668
Author(s):  
Tamar Tadmor ◽  
Yu Zhang ◽  
Robert Dunn ◽  
Seung-uon Shin ◽  
Hyung-Mee Cho ◽  
...  
Keyword(s):  
B Cells ◽  
Tumor Growth ◽  
B Cell ◽  
T Regulatory Cells ◽  
Tumor Response ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Regulatory Cells ◽  
Tumor Bearing ◽  
And Function

Abstract Abstract 3668 Poster Board III-604 Increasing evidence suggests that B lymphocytes play a central role in inhibiting the immune response against certain tumors, but the underlying mechanisms by which B cells facilitate tumor growth are still poorly understood. In this study, we investigated how the presence or absence of B cells affects expansion and function of T- regulatory cells (‘Tregs’) in a murine tumor model (EMT-6). We compared tumor growth, and the number and function of T- regs cells in wild type immune competent mice ( ICM), B cell deficient mice ( BCDM) and /or in BALB-C mice following B- cell depletion induced by injection of anti murine CD20 antibodies (mCD20 Ab, 18B12, mouse IgG1,k, Biogen-IDEC) Mice were either tumor-naïve or implanted with EMT6 mammary adenocarcinoma cells. Absence of B cells as in BCDM completely inhibited tumor growth in the majority of mice, while B cell depletion in normal mice substantially slowed the growth of EMT-6 tumors compared to wild type mice (ICM). Substantial T regs expansion, as defined by CD4+/CD25+/FOXP3+ cells, was evident on day 26 post tumor inoculation in EMT-6 tumor bearing ICM in comparison to the non- tumor bearing mice ( 15.2 +/− 1.2. % and 11.9 +/− 1.1% respectively), isolated from spleen as compared to naïve or tumor bearing BCDM (10.1+/− 0.2% and 10.8+/− 1.2%) The percentage and absolute number of T-regs in the spleen, tumor draining lymph nodes and tumor bed were significantly reduced in the BCDM and/or B cell depleted ICM compared to tumor bearing ICM (10%+/−0.8, 13.9+/− 1.23% and 17+/− 1.3% respectively p<0.01. data from single cell suspensions isolated from spleens on day 20 post tumor inoculation). Similar effects of B cell depletion on the numbers of T-regs were observed in the setting of pre-established EMT6 mammary tumors. In contrast to tumor bearing mice, differences in T-reg number and function were minimal in tumor free B cell deficient or in B cell depleted naïve mice compared to ICM. T-reg function, measured by suppression assay and proliferation assays, was also markedly reduced in tumor bearing BCDM compared to ICM. Combining B cell and T-reg depletion using i.p. injection of anti CD 25 antibody (PC61 or PBS) resulted in similar rates of tumor regression in B cell depleted mice as were seen in BCDM suggesting that the combination of B cells depletion and further depletion of Tregs augmented anti-tumor response. In conclusion, our studies indicate that B cell depletion may play a useful role in augmenting the T cell anti-tumor response, in part due to their effects on T-regulatory cell biology. Disclosures: Dunn: Biogen IDEC: Employment.

CD4+CD25+Foxp3+ T Regulatory Cells Protect Against Murine Antibody-Mediated Transfusion-Related Acute Lung Injury (TRALI)

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2342-2342
Author(s):  
Rick Kapur ◽  
Michael Kim ◽  
Shanjee Shanmugabhavananthan ◽  
Edwin R. Speck ◽  
Rukhsana Aslam ◽  
...  
Keyword(s):  
T Cells ◽  
Cd4 T Cells ◽  
T Regulatory Cells ◽  
Blood Product ◽  
Lung Damage ◽  
Chronic Alcohol ◽  
Regulatory Cells ◽  
Chronic Alcohol Abuse ◽  
Murine Antibody

Abstract Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related fatalities and is characterized by acute respiratory distress following transfusion of blood products. Frequently, donor antibodies present in the transfused blood product are involved, such as anti-human leukocyte antigen (HLA) antibodies or anti-human neutrophil antigen (HNA) antibodies. Several animal models of TRALI have contributed to understanding the pathogenesis which, however, is still incompletely understood. Several cell types have also been suggested to be involved in antibody-mediated TRALI, including neutrophils, endothelial cells and monocytes. Most of the animal models are based on a two-hit paradigm, where the first hit is based on "patient predisposition", such as sepsis or chronic alcohol abuse, while the second hit is delivered in the form of "transfusion factors", such as antibodies present in the transfused blood product. Although most studies have focused on factors contributing to the development of antibody-mediated TRALI, the factors and mechanisms in place to protect against antibody-mediated TRALI have been underexplored. Adoptive transfer of lymphocytes into recipient severe combined immunodeficient (SCID) mice, in which the well-established TRALI inducing anti-MHC class I antibody clone 34-1-2s was injected, was previously shown to rescue TRALI induction by 34-1-2s. Here we describe, using a murine BALB/c antibody-mediated TRALI model based on injection of 34-1-2s, that CD4 T cells, and more specifically, CD4+CD25+Foxp3+ T regulatory cells (Tregs), are responsible for protection against murine antibody-mediated TRALI. Specific in vivo depletion of CD4+ T cells, or targeted in vivo depletion of Tregs, resulted in severe lung damage after 34-1-2s infusion, as determined by increased lung wet-to-dry ratios (a measure for pulmonary edema), generally greater than 5, indicative of severe pulmonary edema. This was accompanied by significant hypothermia, increased values of the neutrophil chemoattractant macrophage inflammatory protein 2 (MIP-2: equivalent of human IL-8), and increased pulmonary neutrophil accumulation, all compared to control groups. In contrast, systematic in vivo depletion of CD8+ T cells, B cells or monocytes, did not result in significant lung damage. Co-depletion of CD4+ T cells together with monocytes rescued the TRALI induction by 34-1-2s, validating the pathogenic role of monocytes in murine antibody-mediated TRALI induction. Based on MIP-2 values and in vitro studies, we suggest that Tregs suppress monocytes in order to prevent antibody-mediated TRALI. Overall, a novel first hit in TRALI induction could be identified in conditions that cause a decrease in Treg number or function, which could also explain the increased risk for human TRALI in cases of chronic alcohol abuse. In addition, therapies aimed at restoring Treg numbers or function may prove to be a novel therapeutic approach in antibody-mediated TRALI. Disclosures No relevant conflicts of interest to declare.

scholarly journals T Regulatory Cells 1 Inhibit a Th2-Specific Response In Vivo

2000 ◽  
Vol 165 (9) ◽  
pp. 4848-4853 ◽  
Author(s):  
Françoise Cottrez ◽  
Steven D. Hurst ◽  
Robert L. Coffman ◽  
Hervé Groux
Keyword(s):  
T Regulatory Cells ◽  
Specific Response ◽  
Regulatory Cells
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